Down-regulation of Twist1 increases the sensitivity of nasopharyngeal carcinoma cell lines HNE1 to taxol
10.3760/cma.j.issn.1673-0860.2009.06.014
- VernacularTitle:降低Twist1表达可以增加鼻咽癌细胞系HNE1对紫杉醇的敏感性
- Author:
Da-Wei MENG
1
;
Ji-Min BAO
;
Yun-Peng MA
;
Zhe LI
;
Su-Jie LI
;
Dan LI
Author Information
1. 辽宁省金秋医院
- Keywords:
Tumor cells,cultured;
Nasopharyngeal neoplasms;
Carcinoma,squamous cell;
Twist transcription factor;
Paclitaxel;
Drug tolerance
- From:
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
2009;44(6):504-508
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate whether down-regulation of Twist1 could change sensitivity of nasopharyngeal carcinoma cell line HNE1 to taxol. Methods HNE1 cells were transfected with the small interfering RNA (siRNA) expression vector pSuppressor-Retro-Si-Twist, containing the short hairpin RNA (shRNA) sequence targeting the Twist gene-coding region by Fugene 6. Positive clones were then selected in Neomycin (400 μg/ml) for 21 days. The low expressions of Twist1 were examined by real-time reverse-transcription polymerase chain reaction (RT-PCR) and Western blot. Drug sensitivity of si-Twist1 HNE1 to taxol was determined by Annexin V-fluorescein isothiocyanate( FITC)/prepidium Iodide(PI) double-labeled flow cytometry and detection of DNA ladder. The Effect of Twist1 inactivation on HNE1 cell proliferation was observed by MTT assay and flow cytometry. Results Annexin V- FITC-PI assay showed that apoptosis ratio was 40.2% in si-Twist HNE1 after treated with 10 ng/ml taxol, significantly higher than that in the control siRNA group 24. 3%. The deference had statistic meaning. After the re-expression of HNE1, apoptosis ratio was 44. 80% ± 4. 80% (x±s) in low Twist1 protein expression group and that was 27. 00%±2. 91% in high expression group. The deference had statistic meaning ( t = 4. 374, P = 0. 049). Real time PCR test revealed apoptosis protein bcl-2 expression in ai-Twist HNE1 was 0. 28±0. 05, significantly lower than that in the control siRNA HNE1 (0. 57 ± 0. 08,t = 6. 710, P = 0. 021 ), nevertheless, significant bax and bcl-XL changes were not observed (t = 2. 000, P = 0. 184 and t = 1. 502, P = 0. 272 ). MTT and FCM showed that down-regulation of Twist1 did not alter cell proliferation rate ( P > 0. 05 ). Conclusions Down-regulation of Twist1 could increase drug sensitivity of nasopharyngeal carcinoma cell line HNE1 to taxol by inducing apoptosis. These results suggested that Twist1 may be a promising treatment target for nasopharyngeal carcinoma therapy.
